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SILIP: a novel stable isotope labeling method for in planta quantitative proteomic analysis
Jennifer E Schaff1, Flaubert Mbeunkui, Kevin Blackburn
1Department of Plant Pathology, Center for the Biology of Nematode Parasitism, North Carolina State University, Raleigh, NC 27695-7253, USA.
The Plant Journal : for Cell and Molecular Biology
|July 31, 2008
Summary
Stable isotope labeling in planta (SILIP) enables accurate in vivo quantitative proteomic analysis in soil-grown tomato plants. This method achieves high 15N incorporation, facilitating plant-microorganism system studies.
Area of Science:
- Plant Biology
- Proteomics
- Metabolomics
Background:
- Metabolic isotope coding for proteomic analysis is typically done in cell culture, limiting in vivo quantitative accuracy.
- Accurate in vivo quantitative analysis is crucial for understanding complex biological systems in intact organisms.
Purpose of the Study:
- To develop a method for stable isotope labeling in planta (SILIP) for quantitative proteomic analysis in soil-grown plants.
- To enable accurate in vivo quantitative proteomic studies in plants, overcoming limitations of cell culture-based methods.
Main Methods:
- Developed SILIP using tomato plants (Solanum lycopersicum) with a 14N/15N isotope coding strategy.
- Plants were grown for 2 months on 14N and 15N-enriched nitrogen sources.
- Proteins were extracted from four distinct tissues and analyzed using LC/MS/MS (DDA) and MS(E).
Main Results:
- Achieved nearly 99% 15N incorporation across various tomato tissues (roots, stems, leaves, flowers), demonstrating uniform labeling.
- Measured 15N incorporation values ranged from 98.2% in leaves to 98.8% in stems.
- SILIP is compatible with root-knot nematode development, enabling simultaneous study of plant-microorganism interactions.
Conclusions:
- SILIP provides a robust and efficient method for in vivo quantitative proteomic analysis in soil-grown plants.
- This technique significantly advances the ability to study plant biological processes and plant-microorganism systems.
- SILIP offers a novel tool for quantitative proteomics in plant science and related fields.
